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Abstract

Cross-neutralization between rabies virus (RABV) and two European bat lyssaviruses (EBLV-1 and -2) was analysed using lentiviral pseudotypes as antigen vectors. Glycoprotein (G-protein) cDNA from RABV challenge virus standard-11 (CVS-11) and EBLV-1 and -2 were cloned and co-expressed with human immunodeficiency virus (HIV) or murine leukemia virus (MLV) gag–pol and packageable green fluorescent protein (GFP) or luciferase reporter genes in human cells. The harvested lentiviral (HIV) vector infected over 40% of baby hamster kidney (BHK) target cells, providing high-titre pseudotype stocks. Tests on blinded antibody-positive (n515) and -negative (n545) sera, predetermined by the fluorescent antibody virus neutralization (FAVN) test approved by the World Health Organization (WHO) and Office International des Epizooties (OIE), revealed that the CVS-11 pseudotype assay had 100% concordance with FAVN and strongly correlated with neutralization titres (r250.89). Cross-neutralization tests using sera from RABVvaccinated humans and animals on pseudotypes with CVS-11, EBLV-1 and EBLV-2 envelopes showed that the relative neutralization titres correlated broadly with the degree of G-protein diversity. Pseudotypes have three major advantages over live-virus neutralization tests: (i) they can be handled in low-biohazard-level laboratories; (ii) the use of reporter genes such as GFP or b-galactosidase will allow the assay to be undertaken at low cost in laboratories worldwide; (iii) each assay requires 10 ml serum. This robust microassay will improve our understanding of the protective humoral immunity that current rabies vaccines confer against emerging lyssaviruses, and will be applicable to surveillance studies, thus helping to control the spread of rabies.

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[1] A supplementary figure and two supplementary tables are available with the online version of this paper see: http://vir.sgmjournals.org/content/89/9/2204/suppl/DC1
[2] These data were presented at the ‘Rabies in the Americas’ (RITA) conference, held in Guanajuato, Mexico in October 2007
[3] This is an open access article published by the Society for General Microbiology under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/).
[4] Citation: Edward Wright, Nigel J. Temperton, Denise A. Marston, Lorraine M. McElhinney, Anthony R. Fooks, and Robin A. Weiss
Investigating antibody neutralization of lyssaviruses using lentiviral pseudotypes: a cross-species comparison J Gen Virol September 2008 89:2204-2213; doi:10.1099/vir.0.2008/000349-0